Ultrasensitive proteome analysis using paramagnetic bead technology

By Christopher S. Hughes, Sophia Foehr, D. Garfield, E. Furlong, Lars M. Steinmetz, Jeroen Krijgsveld. Molecular Systems Biology, 2014

1,098 citations recorded by the source corpus.

In order to obtain a systems‐level understanding of a complex biological system, detailed proteome information is essential. Despite great progress in proteomics technologies, thorough interrogation of the proteome from quantity‐limited biological samples is hampered by inefficiencies during processing. To address these challenges, here we introduce a novel protocol using paramagnetic beads, termed Single‐Pot Solid‐Phase‐enhanced Sample Preparation (SP3). SP3 provides a rapid and unbiased means of proteomic sample preparation in a single tube that facilitates ultrasensitive analysis by outperforming existing protocols in terms of efficiency, scalability, speed, throughput, and flexibility. To illustrate these benefits, characterization of 1,000 HeLa cells and single Drosophila embryos is used to establish that SP3 provides an enhanced platform for profiling proteomes derived from sub‐microgram amounts of material. These data present a first view of developmental stage‐specific proteome dynamics in Drosophila at a single‐embryo resolution, permitting characterization of inter‐individual expression variation. Together, the findings of this work position SP3 as a superior protocol that facilitates exciting new directions in multiple areas of proteomics ranging from developmental biology to clinical applications.

Subfields: Molecular Biology, Spectroscopy

View Paper: https://www.semanticscholar.org/paper/47831fdfc9bc599bf5cf9e96e1c95ea06fd041f8

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Citations in the corpus

A new antibiotic selectively kills Gram-negative pathogens (2019, positive): “After cell lysis, protein aggregates were removed and the remaining soluble proteins were digested according to a modified SP3 protocol 49,50 , as previously described 51 .”
SCoPE-MS: mass spectrometry of single mammalian cells quantifies proteome heterogeneity during cell differentiation (2017, methodology): “These cleanup procedures can result in substantial losses of protein, and colleagues have developed advanced methods, such as SP3 [24] and iST [25], that minimize cleanup losses and allow for quantifying thousands of proteins from samples having just a few micrograms of total protein [23, 26].”